Transfer of cGAMP into Bystander Cells via LRRC8 Volume-Regulated Anion Channels Augments STING-Mediated Interferon Responses and Anti-viral Immunity

Chun Zhou; Xia Chen; Rosa Planells-Cases; Jiachen Chu; Li Wang; Limin Cao; Zhihong Li; Karen I López-Cayuqueo; Yadong Xie; Shiwei Ye; Xiang Wang; Florian Ullrich; Shixin Ma; Yiyuan Fang; Xiaoming Zhang; Zhikang Qian; Xiaozheng Liang; Shi-Qing Cai; Zhengfan Jiang; Dongming Zhou; Qibin Leng; Tsan S Xiao; Ke Lan; Jinbo Yang; Huabin Li; Chao Peng; Zhaozhu Qiu; Thomas J Jentsch; Hui Xiao

doi:10.1016/j.immuni.2020.03.016

Transfer of cGAMP into Bystander Cells via LRRC8 Volume-Regulated Anion Channels Augments STING-Mediated Interferon Responses and Anti-viral Immunity

Immunity. 2020 May 19;52(5):767-781.e6. doi: 10.1016/j.immuni.2020.03.016. Epub 2020 Apr 10.

Authors

Chun Zhou¹, Xia Chen², Rosa Planells-Cases³, Jiachen Chu⁴, Li Wang¹, Limin Cao¹, Zhihong Li⁵, Karen I López-Cayuqueo³, Yadong Xie¹, Shiwei Ye⁶, Xiang Wang⁷, Florian Ullrich³, Shixin Ma¹, Yiyuan Fang⁷, Xiaoming Zhang⁷, Zhikang Qian⁷, Xiaozheng Liang⁷, Shi-Qing Cai⁶, Zhengfan Jiang⁸, Dongming Zhou⁷, Qibin Leng⁹, Tsan S Xiao¹⁰, Ke Lan¹¹, Jinbo Yang¹², Huabin Li¹³, Chao Peng⁵, Zhaozhu Qiu¹⁴, Thomas J Jentsch¹⁵, Hui Xiao¹⁶

Affiliations

¹ The Center for Microbes, Development and Health, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
² The Center for Microbes, Development and Health, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China; College of Life Sciences, Lanzhou University, Lanzhou, Gansu 730000, China.
³ Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) and Max-Delbrück-Centrum für Molekulare Medizin, D-13125 Berlin, Germany.
⁴ Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
⁵ National Facility for Protein Science in Shanghai, Zhangjiang Lab, Shanghai Advanced Research Institute, Chinese Academy of Science, Shanghai 201210, China; Shanghai Science Research Center, Chinese Academy of Sciences, Shanghai 201204, China.
⁶ Institute of Neuroscience and State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
⁷ CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
⁸ Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, School of Life Sciences, Peking University, Peking-Tsinghua Center for Life Sciences, Beijing 100871, China.
⁹ Affiliated Cancer Hospital & Institute of Guangzhou Medical University, State Key Laboratory of Respiratory Diseases, Guangzhou Medical University, Guangzhou, Guangdong 510180, China.
¹⁰ Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
¹¹ State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
¹² College of Life Sciences, Lanzhou University, Lanzhou, Gansu 730000, China.
¹³ Center for Allergic and Inflammatory Diseases & Department of Otolaryngology, Head and Neck Surgery, Affiliated Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai 200031, China.
¹⁴ Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: [email protected].
¹⁵ Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) and Max-Delbrück-Centrum für Molekulare Medizin, D-13125 Berlin, Germany; NeuroCure Cluster of Excellence, Charité University Medicine, D-10117 Berlin, Germany. Electronic address: [email protected].
¹⁶ The Center for Microbes, Development and Health, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: [email protected].

PMID: 32277911
DOI: 10.1016/j.immuni.2020.03.016

Abstract

The enzyme cyclic GMP-AMP synthase (cGAS) senses cytosolic DNA in infected and malignant cells and catalyzes the formation of 2'3'cGMP-AMP (cGAMP), which in turn triggers interferon (IFN) production via the STING pathway. Here, we examined the contribution of anion channels to cGAMP transfer and anti-viral defense. A candidate screen revealed that inhibition of volume-regulated anion channels (VRACs) increased propagation of the DNA virus HSV-1 but not the RNA virus VSV. Chemical blockade or genetic ablation of LRRC8A/SWELL1, a VRAC subunit, resulted in defective IFN responses to HSV-1. Biochemical and electrophysiological analyses revealed that LRRC8A/LRRC8E-containing VRACs transport cGAMP and cyclic dinucleotides across the plasma membrane. Enhancing VRAC activity by hypotonic cell swelling, cisplatin, GTPγS, or the cytokines TNF or interleukin-1 increased STING-dependent IFN response to extracellular but not intracellular cGAMP. Lrrc8e^-/- mice exhibited impaired IFN responses and compromised immunity to HSV-1. Our findings suggest that cell-to-cell transmission of cGAMP via LRRC8/VRAC channels is central to effective anti-viral immunity.

Keywords: 3'3'cGAMP; 3'3'cGMP-AMP; MCMV; VSOAC; VSOR; adenovirus; c-di-AMP; c-di-GMP; cdA; cdG; herpesvirus; innate immunity; retrovirus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antiviral Agents / immunology
Antiviral Agents / metabolism
Bystander Effect
Cell Line
Cells, Cultured
Embryo, Mammalian / cytology
Embryo, Mammalian / metabolism
Fibroblasts / cytology
Fibroblasts / immunology*
Fibroblasts / metabolism
HeLa Cells
Herpes Simplex / immunology
Herpes Simplex / virology
Herpesvirus 1, Human / immunology
Herpesvirus 1, Human / physiology
Humans
Interferons / immunology*
Interferons / metabolism
Membrane Proteins / genetics
Membrane Proteins / immunology*
Membrane Proteins / metabolism
Mice, Inbred C57BL
Mice, Knockout
Nucleotides, Cyclic / immunology*
Nucleotides, Cyclic / metabolism
Nucleotidyltransferases / genetics
Nucleotidyltransferases / immunology
Nucleotidyltransferases / metabolism
Voltage-Dependent Anion Channels / immunology*
Voltage-Dependent Anion Channels / metabolism

Substances

Antiviral Agents
LRRC8A protein, mouse
Membrane Proteins
Nucleotides, Cyclic
Sting1 protein, mouse
Voltage-Dependent Anion Channels
cyclic guanosine monophosphate-adenosine monophosphate
Interferons
Nucleotidyltransferases
cGAS protein, mouse